探討膽囊收縮素受體A在人類大腸癌細胞的表現量以及與細胞移動和侵襲能力的關聯性

Investigating expression of cholecystokinin receptor A in human colon carcinoma cells and its linking to cell motility and invasiveness

林建賢(Chien-Hsien Lin)

指導教授 ： 高紹軒

中山醫學大學/醫學院/生化暨生物科技研究所/碩士(2016年)

摘要

大腸癌長年位居國內十大癌症的前三名，目前穩坐於榜首位置，在美國癌症協會的統計資料中，無論男性或女性的癌症發生率排名第三名；膽囊收縮素受體的功用主要與腸胃道的膽囊收縮素和胃泌素結合，調節飲食攝取和身體能量代謝，膽囊收縮素受體在癌症發展過程也扮演著一關鍵角色，尤其是發炎性腸胃道癌症，包括大腸癌。利用傷口癒合和Transwell實驗研究人類的4種大腸癌細胞，觀察到其中DLD-1細胞和HT-29細胞擁有較強的爬行和侵襲能力，相較於Caco-2和LoVo細胞；接著研究這4株細胞內的膽囊收縮素受體A和B表現量，由Western blot結果顯示LoVo細胞中含有最多的膽囊收縮素受體A和B，其次是DLD-1細胞。其中DLD-1細胞同時具有高遷移性與侵襲能力，因此進一步利用siRNA來關閉膽囊收縮素受體A的表現，並藉由傷口癒合和Transwell實驗來分析膽囊收縮素受體A的表現對DLD-1細胞遷移性與侵襲能力之影響。結果發現使用siRNA處理過的DLD-1細胞移動和侵襲的能力會顯著下降，推測膽囊收縮素受體A的表現可能促使大腸腫瘤由原位癌演變成轉移性大腸癌；後續研究將再針對膽囊收縮素受體A與細胞移動與大腸癌移轉之分子機制進行更深入之分析，以及探討是否可以發展新型的大腸癌治療標的。

關鍵字

膽囊收縮素受體A ； 大腸癌 ； 細胞移動 ； 細胞侵襲能力

並列摘要

The Colon cancer is the highest prevalence cancer in Taiwan and the third high prevalence cancer in United States. Cholecystokinin (CCK) receptors are widely expressed in digestive tract and exert their physiological functions such as regulating dietary intake and body energy metabolism after binding with their ligands CCK and gastrin secreted from gastrointestinal track. Recent studies have shown that CCK receptor also plays a key role in the cancer development process, particularly in inflammatory gastrointestinal cancers such as colon cancer. However, whether expression of CCK receptors involve in motility of colon carcinoma cells remain sketchy. Therefore, we aimed to investigate the expression of CCK type A and type B receptor (CCKAR and CCKBR) in different colon carcinoma cell line and the association between expression of CCK receptors and cell motility. Four colon cancer cell lines Caco-2, DLD-1, Lovo and HT-29 were used for examination and cell motility were analyzed by using wound healing and transwell assay. Expression of CCK receptors was determined by Western blot. Our results showed that DLD-1 and HT-29 cell showed the highest motility in wound healing assay and transwell assay, respectively. Western blot analysis revealed that both CCKAR and CCKBR was highly expressed in Lovo cell and showed relative low expression in HT-29 cell. The similar expression level of CCKAR and CCKBR was also observed by using RT-PCR. Knock down expression of CCKAR by using specific siRNA significantly reduced the cell motility of DLD-1 cell in wound healing and transwell assay. In conclusion, our findings indicate that CCKAR expression may associate with cell motility of colon cancer cell and play an important role in promoting cell motility of colon cancer cell. These results suggest that CCKAR might be a potential target for developing colon cancer treatment via inhibition of tumor metastasis.

並列關鍵字

Cholecystokinin receptor A ； Colon cancer ； Cell motility ； Cell invasiveness

參考文獻

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